The cyathins are a unique group of diterpenoids produced by the birds nest fungi <i>Cyathus helenae</i>, <i>C. africanus</i>, and <i>C. earlei</i>. Several of the cyathins show strong antibiotic activity. More recently, several fungal metabolites with structures closely related to those of the cyathins have been found to be potent inducers of nerve growth factor (NGF) synthesis. The structural complexity and the exciting biological activity of the cyathane family of diterpenes have prompted our efforts to develop an efficient and general synthetic approach.<p>To date, there have been seven total syntheses and six partial syntheses of cyathins. Most of these syntheses target allocyathin B2, which does not contain the common 6-7 trans ring fusion or the hydroxyl group within the seven member ring. Modifications of these routes to provide targets with these features have not been demonstrated and may be challenging. We have developed a concise asymmetric synthesis of cyathin A3, based on the enantioselective Diels-Alder reaction of quinone (106) and diene (105). Because the transformations of cyathin A3 into other cyathins are well documented, this synthesis provides a general approach to the cyathane diterpene family. In Section 2.2, the enantioselective Diels-Alder reaction of quinone 106 and diene 105 is presented. This reaction is effectively catalyzed by a carefully prepared Mikami catalyst. It was carried out on a preparative scale to give the chiral building block 108. The absolute configuration of the Diels-Alder adduct 108 was determined by NMR and X-ray analysis.<p>In Sections 2.3-5, the enantioselective total synthesis of (-)-cyathin A3 is described. This approach features the successful oxymercuration ring opening, a newly developed in situ configuration inversion, a much improved intramolecular aldol reaction and a radical cyclization. Now envisioned in our laboratory is the development of a new access to cyathin A4, which is surmised to be possible via the intermediate prepared in this synthesis.

The cyathins are a unique group of diterpenoids produced by the birds nest fungi <i>Cyathus helenae</i>, <i>C. africanus</i>, and <i>C. earlei</i>. Several of the cyathins show strong antibiotic activity. More recently, several fungal metabolites with structures closely related to those of the cyathins have been found to be potent inducers of nerve growth factor (NGF) synthesis. The structural complexity and the exciting biological activity of the cyathane family of diterpenes have prompted our efforts to develop an efficient and general synthetic approach.<p>To date, there have been seven total syntheses and six partial syntheses of cyathins. Most of these syntheses target allocyathin B2, which does not contain the common 6-7 trans ring fusion or the hydroxyl group within the seven member ring. Modifications of these routes to provide targets with these features have not been demonstrated and may be challenging. We have developed a concise asymmetric synthesis of cyathin A3, based on the enantioselective Diels-Alder reaction of quinone (106) and diene (105). Because the transformations of cyathin A3 into other cyathins are well documented, this synthesis provides a general approach to the cyathane diterpene family. In Section 2.2, the enantioselective Diels-Alder reaction of quinone 106 and diene 105 is presented. This reaction is effectively catalyzed by a carefully prepared Mikami catalyst. It was carried out on a preparative scale to give the chiral building block 108. The absolute configuration of the Diels-Alder adduct 108 was determined by NMR and X-ray analysis.<p>In Sections 2.3-5, the enantioselective total synthesis of (-)-cyathin A3 is described. This approach features the successful oxymercuration ring opening, a newly developed in situ configuration inversion, a much improved intramolecular aldol reaction and a radical cyclization. Now envisioned in our laboratory is the development of a new access to cyathin A4, which is surmised to be possible via the intermediate prepared in this synthesis.

Orientador: Valéria Maia Merzel / Dissertação (mestrado) - Universidade Estadual de Campinas, Insituto de Biologia / Made available in DSpace on 2018-08-16T20:58:52Z (GMT). No. of bitstreams: 1 Silva_TiagoRodriguese_M.pdf: 5632148 bytes, checksum: 82526f541aaf1c9b32cf5fbca6bc03aa (MD5) Previous issue date: 2010 / Resumo: Estudos realizados em reservatórios de petróleo têm evidenciado que parte da microbiota associada a este tipo de ambiente é representada por bactérias e arqueias de distribuição geográfica bastante ampla e que diversos destes organismos têm potencial para transformar compostos orgânicos e inorgânicos, atuando na interface óleo-água dos reservatórios. A investigação de micro-organismos com potencial para biodeterioração, biodegradação e biocorrosão encontrados em depósitos petrolíferos é de grande importância, uma vez que estes organismos podem estar relacionados com a perda da qualidade do petróleo nos reservatórios e etapas subseqüentes de exploração. Este estudo teve como finalidade comparar a microbiota presente em amostras de óleo de dois poços de petróleo terrestres da Bacia Potiguar (RN), identificados como GMR75 (poço biodegradado) e PTS1 (poço não-biodegradado). As comunidades microbianas foram estudadas usando técnicas de cultivo (enriquecimentos microbianos e isolamento) e independentes de cultivo (construção de bibliotecas de genes RNAr 16S). Os micro-organismos cultivados de ambos os poços mostraram-se afiliados aos filos Actinobacteria, Firmicutes e Proteobacteria. As bibliotecas de gene RNAr 16S foram construídas a partir de DNA total extraído do petróleo bruto. Ambas as bibliotecas de bactérias revelaram uma grande diversidade, com 8 filos diferentes para o poço GMR75, Actinobacteria, Bacteroidetes, Deferribacteres, Spirochaetes, Firmicutes, Proteobacteria, Thermotoga e Synergistetes, e 5 filos para o poço PTS1, Actinobacteria, Chloroflexi, Firmicutes, Proteobacteria e Thermotogae. A biblioteca de genes RNAr 16S de arqueias só foi obtida para o poço GMR75 e todos os clones encontrados mostraram-se relacionados a membros da ordem Methanobacteriales. Os resultados de diversidade sugerem que a metanogênese é o processo terminal dominante no poço, o que indica uma biodegradação anaeróbia. A comparação dos estudos dependente e independente de cultivo mostrou que alguns gêneros, como Janibacter, Georgenia, Saccharopolyspora, Tessaracoccus, Brevundimonas e Brachymonas não foram encontradas na abordagem independente de cultivo, sugerindo que mais clones devam ser seqüenciados para cobrir toda a diversidade presente na amostra. Nossa hipótese de que poderia haver algum agente antimicrobiano inibindo o crescimento de bactérias degradadoras de hidrocarbonetos no poço não-biodegradado não foi confirmada. No entanto, durante os testes realizados, uma bactéria, Bacillus pumilus, isolada em estudos anteriores de reservatórios da Bacia de Campos, apresentou resultados positivos de inibição para todas as linhagens testadas como indicadoras, e os testes de caracterização do composto revelaram ser este um diterpeno da classe das Ciatinas. / Abstract: Recent studies from oil fields have shown that microbial diversity is represented by bacteria and archaea of wide distribution, and that many of these organisms have potential to metabolize organic and inorganic compounds. The potential of biodeterioration, biodegradation and biocorrosion by microorganisms in oil industry is of great relevance, since these organisms may be related with the loss of petroleum quality and further exploration steps. The aim of the present study was to compare the microbial communities present in two samples from terrestrial oil fields from Potiguar basin (RN - Brazil), identified as GMR75 (biodegraded oil) and PTS1 (non-biodegraded oil). Microbial communities were investigated using cultivation (microbial enrichments and isolation) and molecular approaches (16S rRNA gene clone libraries). The cultivated microorganisms recovered from both oil-fields were affiliated with the phyla Actinobacteria, Firmicutes and Proteobacteria. The 16S rRNA gene clone libraries were constructed from metagenomic DNA obtained from crudeoil. Both bacterial libraries revealed a great diversity, encompassing representatives of 8 different phyla for GMR75, Actinobacteria, Bacteroidetes, Deferribacteres, Spirochaetes, Firmicutes, Proteobacteria, Thermotogae and Synergistetes, and of 5 different phyla, Actinobacteria, Chloroflexi, Firmicutes, Proteobacteria and Thermotoga, for PTS1. The archaeal 16S rRNA clone library was obtained only for GMR75 oil and all phylotypes were affiliated with order Methanobacteriales. Diversity resuts suggest that methanogenesis is the dominant terminal process in GMR75 reservoir, driven by anaerobic biodegradation. The cross-evaluation of culture-dependent and independent techniques indicates that some bacterial genera, such as Janibacter, Georgenia, Saccharopolyspora, Tessaracoccus, Brevundimonas and Brachymonas, were not found using the the 16S rRNA clone library approach, suggesting that additional clones should be sequenced in order to cover diversity present in the sample. Our hypothesis that biodegrading bacterial populations could be inhibited by antimicrobialproducing microorganisms in the non biodegraded oil field (PTS1) was not confirmed. However, one Bacillus pumilus strain, previously isolated from Campos Basin reservoirs, showed positive results in inhibitory tests for all indicator strains. Chemical analyses allowed us to identify the compound as a diterpen from the Cyathin class. / Mestrado / Genetica de Microorganismos / Mestre em Genética e Biologia Molecular